Novel proline substituted cyclosporin analogues

ABSTRACT

The present invention provides novel proline substituted cyclosporinanalogue compounds, pharmaceutical compositions comprising these compounds and methods of using these compounds for the treatment of disorders and diseases, including immune disorders, inflammatory disorders and viral infections.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.61/224,343, filed on Jul. 9, 2009. The entire teachings of the aboveapplication are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to novel cyclosporin analogues,compositions containing them, processes for their preparation,intermediates in their synthesis, and their use as therapeutics forprevention of organ transplantation rejection, the treatment of immunedisorders and inflammation, and treatment of viral (particularlyheptitis C viral) infection.

BACKGROUND OF THE INVENTION

Cyclosporin A (CsA), a neutral cyclic undecapeptide isolated from thefungus Tolypocladium injlaturn and currently marketed as Neoral andsandimmunem (Novartis, Basel, Switzerland), has been widely used for theprevention of organ transplant rejection. The molecular basis for theimmunosuppressant activity of cyclosporin A and cyclosporin analoguesbegins with the passive diffusion of the cyclosporin (Cs) molecule intothe cell, followed by binding to its intracellular receptor, cyclophilinA (CypA). CypA belongs to a family of proteins that catalyze cis-transpeptidyl-prolyl isomerization, i.e., PPIase, a rate-limiting step inprotein folding. CsA and other cyclosporin analogues bind to the activesite of CypA. However, immunosuppression is not believed to be due tothe inhibition of CypA PPIase activity. The target of the CsA-CypAcomplex is a Ca²⁺-calmodulin-dependent serine-threonine-specific proteinphosphatase, calcineurin. In T-cells responding to antigen presentation,an increase in intracellular Ca²⁺ activates calcineurin, whichsubsequently dephosphorylates the transcription factor called thenuclear factor of activated T-cells (“NFAT”). Dephosphorylated NFATundergoes a molecular change, e.g., homodimerization that allows it tocross into the nucleus, and promotes the expression of T-cell activationgenes. CsA and other immunosuppressive cyclosporin derivatives inhibitcalcineurin which results in the inhibition of expression of cytokinegenes, e.g., interleukin-2 (IL-2) that promotes T-cell activation andproliferation, i.e., immunosuppressive activity.

Since the original discovery of cyclosporin, a wide variety of naturallyoccurring cyclosporins have been isolated and identified, and manyfurther nonnatural cyclosporins have been prepared by total- orsemi-synthetic means or by the application of modified culturetechniques. The class comprised by the cyclosporins is thus nowsubstantial and includes, for example, the naturally occurringcyclosporins A through Z [cf., Traber et al.; 1, Helv. Chim. Acta, 60,1247-1255 (1977); Traber et al.; 2, Helv. Chim. Acta, 65, 1655-1667(1982); Kobel et al.; Europ. J. Applied Microbiology and Biotechnology,14, 273-240 (1982); and von Wartburg et al.; Progress in Allergy, 38,28-45 (1986)], as well as various non-natural cyclosporin derivativesand artificial or synthetic cyclosporin derivatives and artificial orsynthetic cyclosporins including dihydrocyclosporins [in which the-MeBmt-residue is saturated by hydrogenation]; derivatized cyclosporins(e.g., in which the 3′-O-atom of the -MeBmt- residue is acylated or afurther substituent is introduced at the a-carbon atom of the sarcosylresidue at the 3-position); and cyclosporins in which variant aminoacids are incorporated at specific positions within the peptidesequence, e.g. employing the total synthetic method for the productionof cyclosporins developed by R. Wenger—see e.g. Traber et al., 1; Traberet al., 2; and Kobel et al., loc cit. U.S. Pat. Nos. 4,108,985,4,220,641, 4,288,431, 4,554,351, 4,396,542 and 4,798,823; EuropeanPatent Publication Nos. 34,567A, 56,782A, 300,784A and 300,785;International Patent Publication No. WO 86/02080 and UK PatentPublication Nos. 2,206,119 and 2,207,678; Wenger 1, Transpl. Proc., 15Suppl. 1:2230 (1983); Wenger 2, Angew. Chem. Int. Ed. 24 77 (1985) andWenger 3, Progress in the Chemistry of Organic Natural Products, 50, 123(1986). Several synthetic modifications of the -MeBmt- residue residingat position 1 of the cyclosporin undecapeptide have been describedincluding: Park et al., Tetrahedron Lett. 1989, 30, 4215-4218; U.S. Pat.Nos. 5,239,037, 5,293,057; U.S. Publication Nos. US20020142946,US20030087813, and US20030104992 assigned to Enanta Pharmaceuticals,Inc.; PCT Publication Nos. WO99/18120 and WO03/033526 both assigned toIsotechnika; and U.S. Pat. Nos. 4,384,996, 4,771,122, 5,284,826, and5,525,590 assigned to Sandoz.

The compound cyclosporin (cyclosporinA or CsA) has found wide use sinceits introduction in the fields of organ transplantation andimmunomodulation, and has brought about a significant increase in thesuccess rate for transplantation procedures. Undesired side effectsassociated with cyclosporin, however, such as nephrotoxicity, have ledto a continued search for immunosuppressant compounds having improved,efficacy and safety.

Side effects with systemic CsA include increase in diastolic bloodpressure and decrease in renal function. Other side effects includehepatic dysfunction, hypertrichosis, tremor, gingival hyperplasia andparaesthsia. The systemic toxicity of CsA limits its use for thetreatment of certain diseases. Accordingly, a need exists for compoundswhich exhibit immunosuppressive activity while not producing systemictoxicity.

Cyclosporin A and certain derivatives have been reported as havinganti-HCV activity, see Watashi et al., Hepatology, 2003, Volume 38, pp1282-1288, Nakagawa et al., Biochem. Biophys. Res. Commun. 2004, Volume3 13, pp 42-7, and Shimotohno and K. Watashi, 2004 American TransplantCongress, Abstract No. 648 (American Journal of Transplantation 2004,Volume 4, Issue s8, Pages 1-653). The authors of the Nakagawa et al.paper state that certain chaperone activities, such as those ofcyclophilins, may be crucial for the processing and maturation of theviral proteins and for viral replication. Cyclosporin derivatives havinganti-HCV activity are known from International Publication Nos. WO2005/021028, WO 2006/039668, WO 2006/038088, WO 2006/039688, WO2007/112352, WO 2007/112357, WO 2007/112345 and WO 2007/041631.

A subsequent controlled clinical trial showed that a combination ofcyclosporin A with interferon α2b is more effective than interferonmonotherapy, especially in patients with high viral loads (Inoue et al.,“Combined Interferon α2b and Cyclosporin A in the Treatment of ChronicHepatitis C: Controlled Trial,” J. Gastroenterol. 38:567-572 (2003)).

PCT International Patent Publication No. WO 2006/005610 recentlydescribed the use of a combination of cyclosporin A and pegylatedinterferon for treating hepatitis C viral infection. In addition, PCTInternational Patent Publication No. WO 2005/021028 relates to the useof non-immunosuppressive cyclosporine for treatment of HCV disorders.Also, Paeshuyse et al., “Potent and Selective Inhibition of Hepatitis CVirus Replication by the Non-Immunosuppressive Cyclosporin AnalogueDEBIO-025,” Antiviral Research 65(3):A41 (2005) recently publishedresults for a non-immunosuppressive cyclosporin analogue, DEBIO-025,that exhibited potent and selective inhibition of hepatitis C virusreplication. Debio-025 does possess potent binding affinity forcyclophilin A.

SUMMARY OF THE INVENTION

The present invention relates to novel cyclosporin analogues representedherein below, pharmaceutical compositions comprising such compounds,methods for the treatment of viral (particularly hepatitis C viral)infection in a subject in need of such therapy with said compounds andmethods of treatment for prevention of organ transplantation rejection,the treatment of immune disorders and inflammation.

In its principal embodiment, the present invention provides a compoundof formula (I);

where:

A is

where, R₁ is selected from;

-   -   a) R₁₁, where R₁₁ is selected from;        -   1) Hydrogen;        -   2) Deuterium;        -   3) C₁-C₈ alkyl;        -   4) Substituted C₁-C₈ alkyl;        -   5) C₂-C₈ alkenyl;        -   6) Substituted C₂-C₈ alkenyl;        -   7) C₂-C₈ alkynyl;        -   8) Substituted C₂-C₈ alkynyl;        -   9) C₃-C₁₂ cycloalkyl;        -   10) Substituted C₃-C₁₂ cycloalkyl;        -   11) Aryl;        -   12) Substituted aryl;        -   13) Heterocycloalkyl;        -   14) Substituted heterocycloalkyl;        -   15) Heteroaryl; and        -   16) Substituted heteroaryl;    -   b) —C(O)OR₁₁, where R₁₁ is as previously defined;    -   c) —C(O)R₁₁, where R₁₁ is as previously defined;    -   d) —C(O)OCH₂—V—R₁₂, where V is —O— or —S— and R₁₂ is selected        from;        -   1) C₁-C₈ alkyl;        -   2) Substituted C₁-C₈ alkyl;        -   3) C₂-C₈ alkenyl;        -   4) Substituted C₂-C₈ alkenyl;        -   5) C₂-C₈ alkynyl;        -   6) Substituted C₂-C₈ alkynyl;        -   7) C₃-C₁₂ cycloalkyl;        -   8) Substituted C₃-C₁₂ cycloalkyl;        -   9) Aryl;        -   10) Substituted aryl;        -   11) Heterocycloalkyl;        -   12) Substituted heterocycloalkyl;        -   13) Heteroaryl; and        -   14) Substituted heteroaryl;    -   e) —C(O)N(R₁₃)(R₁₄), where R₁₃ and R₁₄ are independently        selected from R₁₁ and R₁₁ is as previously defined or R₁₃ and        R₁₄ combined together with the N which attached to is        substituted or unsubstituted heterocycloalkyl;    -   f) —C(O)SR₁₁, where R₁₁ is as previously defined;    -   g) —C(S)OR₁₁, where R₁₁ is as previously defined;    -   h) —C(O)OCH₂OC(O)R₁₂, where R₁₂ is as previously defined;    -   i) —C(S)SR₁₁, where R₁₁ is as previously defined; and    -   j) R₁₅, where R₁₅ is selected from;        -   1) -M-R₁₁, where R₁₁ is as previously defined and M is            selected from;            -   i. C₁-C₈ alkyl;            -   ii. Substituted C₁-C₈ alkyl;            -   iii. C₂-C₈ alkenyl;            -   iv. Substituted C₂-C₈ alkenyl;            -   v. C₂-C₈ alkynyl;            -   vi. Substituted C₂-C₈ alkynyl;            -   vii. C₃-C₁₂ cycloalkyl; and            -   viii. Substituted C₃-C₁₂ cycloalkyl;        -   2) -M-NR₁₃R₁₄, where R₁₃, R₁₄ and M are as previously            defined;        -   3) -M-S(O)_(m)R₁₁, where m=0, 1, or 2; M and R₁₁ are as            previously defined;        -   4) -M-OR₁₁, where M and R₁₁ are as previously defined;        -   5) -M-C(O)R₁₁, where M and R₁₁ are as previously defined;        -   6) -M-OC(O)R₁₂, where M and R₁₂ are as previously defined;        -   7) -M-OC(O)OR₁₂, where M and R₁₂ are as previously defined;        -   8) -M-NR₁₇C(O)R₁₂, where R₁₇ is R₁₁, M and R₁₂ are as            previously defined;        -   9) -MNR₁₇C(O)OR₁₂, where R₁₇, M and R₁₂ are as previously            defined;        -   10) -M-C(O)NR₁₃R₁₄, where R₁₃, M and R₁₄ are as previously            defined;        -   11) -M-C(O)N(R₁₇)—OR₁₁, where R₁₇, M and R₁₁ are as            previously defined;        -   12) -M-OC(O)NR₁₃R₁₄, where R₁₃, M and R₁₄ are as previously            defined;        -   13) -M-NR₁₇C(O)NR₁₆R₁₁, where M, R₁₃, R₁₇ and R₁₄ are as            previously defined;        -   14) -M-C(S)SR₁₁, where M and R₁₁ are as previously defined;        -   15) -M-OC(S)SR₁₂, where M and R₁₂ are as previously defined;        -   16) -M-NR₁₇C(O)SR₁₂, where M, R₁₇ and R₁₂ are as previously            defined;        -   17) -M-SC(O)NR₁₃R₁₄, where M, R₁₃ and R₁₄ are as previously            defined;        -   18) -M-CH═N—OR₁₁, where M and R₁₁ are as previously defined;            and        -   19) -M-CH═N—NR₁₃R₁₄, where M, R₁₃ and R₁₄ are as previously            defined;            B is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;            X is OR₁ or SR₁, where R₁ is as previously defined;            W is absent, —O—, or —S(O)_(m)—, where m=0, 1, or 2;            R₃ is R₁, where R₁ is as previously defined;            R_(4N) is selected from methyl, ethyl, allyl and propyl;            R₄ is —(CH₂)n₁-C(R₄₁)(R₄₂)—W₁—R₁, where n₁=0, 1 or 2; W₁ is            absent, —O—, or —S(O)_(m), where m=0, 1 or 2; R₄₁ and R₄₂            are independently selected from: hydrogen, methyl, ethyl,            allyl, propyl and isopropyl; and R₁ is as previously            defined.

In another embodiment, the present invention provides the use ofcyclosporin analogs for the treatment of, with or without the concurrentuse of other drugs, organ transplantation rejections, immune disorders,and inflammation including, but not limited to, indications such asrheumatoid arthritis, psoriasis, inflammatory bowel diseases, chronicobstructive pulmonary disease, allergic rhinitis, and asthma.

In yet another embodiment, the present invention provides apharmaceutical composition comprising a therapeutically effective amountof a compound or combination of compounds of the present invention, or apharmaceutically acceptable salt form, prodrug, salt of a prodrug,stereoisomer, tautomer, solvate, or combination thereof, in combinationwith a pharmaceutically acceptable carrier or excipient.

In still another embodiment, the present invention provides a method ofinhibiting the replication of an RNA-containing virus comprisingcontacting said virus with a therapeutically effective amount of acompound or a combination of compounds of the present invention, or apharmaceutically acceptable salt, prodrug, salt of a pro drug,stereoisomer, tautomer, solvate, or combination thereof. Particularly,this invention is directed to methods of inhibiting the replication ofhepatitis C virus.

Yet another embodiment, the present invention provides a method oftreating or preventing infection caused by an RNA-containing viruscomprising administering to a patient in need of such treatment atherapeutically effective amount of a compound or combination ofcompounds of the present invention, or a pharmaceutically acceptablesalt form, prodrug, salt of a prodrug, stereoisomer, or tautomer,solvate, or combination thereof. Particularly, this invention isdirected to methods of treating or preventing infection caused byhepatitis C virus.

Yet another embodiment of the present invention provides the use of acompound or combination of compounds of the present invention, or atherapeutically acceptable salt form, prodrug, salt of a prodrug,stereoisomer or tautomer, solvate, or combination thereof, as definedhereinafter, in the preparation of a medicament for the treatment orprevention of infection caused by RNA-containing virus, specificallyhepatitis C virus (HCV).

DETAILED DESCRIPTION OF THE INVENTION

In a first embodiment of the present invention is a compound of formula(I) as illustrated above, or a pharmaceutically acceptable salt, esteror prodrug thereof.

Representative subgenera of the present invention include:

Compounds of formula I which are represented by the formula (II):

wherein R₃, R₄, R_(4N) and W are as defined for formula I and

represents a single bond or a double bond.

Compounds of formula I which are represented by the formula (III):

wherein R₃, R₄ and W are as defined for formula I.

Compounds or formula I which are represented by the formula (IV):

wherein R₃, R₄ and W are as defined for formula I.Representative species of the present invention include:Example 1: Compound of formula IV: R₄=isobutyl, W is absent, and R₃=H;Example 2: Compound of formula IV: R₄=isobutyl, W=O, and R₃=H;Example 3: Compound of formula IV: R₄=isobutyl, W=O, and R₃=Me;Example 4: Compound of formula IV: R₄=isobutyl, W=O, and R₃=Ac;Example 5: Compound of formula IV: R₄=isobutyl, W=O, and R₃=allyl;Example 6: Compound of formula IV: R₄=isobutyl, W=O, and R₃=—C(O)NH₂;Example compounds 7-24 of the formula IV, wherein W, R₄ and R₃ aredelineated for each example in Table I.

TABLE I Ex- am- ple W R₄ R₃ 7 O

8 O

9 O

10 O

11 O

12 O

13 O

14 O

15 O

16 O

17 O

18 O

19 O

20 O

21 O

22 O

23 O

24 O

Example 25: Compound of formula IV: R₄=isopropyl, W=absent, and R₃=H.Example compounds 25-185 of the formula IV, wherein W, R₄ and R₃ aredelineated for each example in Table II.

TABLE II Example W R₄ R₃ 25 absent

H 26 O

H 27 O

Ac 28 O

allyl 29 O

30 O

31 O

32 O

33 O

34 O

35 O

36 O

37 O

38 O

39 O

40 O

41 O

42 O

43 O

44 O

45 O

46 O

47 O

48 absent

H 49 O

H 50 O

Ac 51 O

allyl 52 O

53 O

54 O

55 O

56 O

57 O

58 O

59 O

60 O

61 O

62 O

63 O

64 O

65 O

66 O

67 O

68 O

69 O

70 O

71 absent

H 72 O

H 73 O

Ac 74 O

allyl 75 O

76 O

77 O

78 O

79 O

80 O

81 O

82 O

83 O

84 O

85 O

86 O

87 O

88 O

89 O

90 O

91 O

92 O

93 O

94 absent

H 95 O

H 96 O

Ac 97 O

allyl 98 O

99 O

100 O

101 O

102 O

103 O

104 O

105 O

106 O

107 O

108 O

109 O

110 O

111 O

112 O

113 O

114 O

115 O

116 O

117 absent

H 118 O

H 119 O

Ac 120 O

allyl 121 O

122 O

123 O

124 O

125 O

126 O

127 O

128 O

129 O

130 O

131 O

132 O

133 O

134 O

135 O

136 O

137 O

138 O

139 O

140 absent

H 141 O

H 142 O

Ac 143 O

allyl 144 O

145 O

146 O

147 O

148 O

149 O

150 O

151 O

152 O

153 O

154 O

155 O

156 O

157 O

158 O

159 O

160 O

161 O

162 O

163 absent

H 164 O

H 165 O

Ac 166 O

allyl 167 O

168 O

169 O

170 O

171 O

172 O

173 O

174 O

175 O

176 O

177 O

178 O

179 O

180 O

181 O

182 O

183 O

184 O

185 O

A further embodiment of the present invention includes pharmaceuticalcompositions comprising any single compound delineated herein, or apharmaceutically acceptable salt, ester, solvate, or prodrug thereof,with a pharmaceutically acceptable carrier or excipient.

Yet another embodiment of the present invention is a pharmaceuticalcomposition comprising a combination of two or more compounds delineatedherein, or a pharmaceutically acceptable salt, ester, solvate, orprodrug thereof, with a pharmaceutically acceptable carrier orexcipient.

As immunosuppressants, the compounds of the present invention are usefulwhen administered for the prevention of immune-mediated tissue or organgraft rejection. Examples of transplanted tissues and organs whichsuffer from these effects are heart, kidney, liver, medulla ossium,skin, cornea, lung, pancreas, intestinum tenue, limb, muscle, nervus,duodenum, small-bowel, pancreatic-islet-cell, and the like; as well asgraft-versus-host diseases brought about by medulla ossiumtransplantation. The regulation of the immune response by the compoundsof the invention would also find utility in the treatment of autoimmunediseases, such as rheumatoid arthritis, systemic lupus erythematosis,hyperimmunoglobulin E, Hashimoto's thyroiditis, multiple sclerosis,progressive systemic sclerosis, myasthenia gravis, type I diabetes,uveitis, allergic encephalomyelitis, glomerulonephritis, and the like;and further infectious diseases caused by pathogenic microorganisms,such as HIV. In the particular cases of HIV-1, HIV-2 and relatedretroviral strains, inhibition of T-cell mitosis would suppress thereplication of the virus, since the virus relies upon the host T-cell'sproliferative functions to replicate.

Further uses include the treatment and prophylaxis of inflammatory andhyperproliferative skin diseases and cutaneous manifestations ofimmunologically mediated illnesses, such as psoriasis, atopicaldermatitis, contact dermatitis and further eczematous dermatitises,seborrhoeis dermatitis, Lichen planus, Pemphigus, bullous pemphigoid,Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas,cutaneous eosinophilias, Lupus erythematosus, acne and Alopecia greata;various eye diseases (autoimmune and otherwise) such askeratoconjunctivitis, vernal conjunctivitis, keratitis, herpetickeratitis, conical cornea, dystrophia epithelialis corneae, cornealleukoma, ocular pemphigus, Mooren's ulcer, Scleritis, Graves'ophthalmopathy, Vogt-Koyanagi-Harada syndrome, sarcoidosis, multiplemyeloma, etc.; obstructive airway diseases, which includes conditionssuch as chronic obstructive pulmonary disease (COPD), asthma (forexample, bronchial asthma, allergic asthma, intrinsic asthma, extrinsicasthma and dust asthma), particularly chronic or inveterate asthma (forexample, late asthma and airway hyperresponsiveness), bronchitis,allergic rhinitis and the like; inflammation of mucosa and blood vesselssuch as gastric ulcers, vascular damage caused by ischemic diseases andthrombosis. Moreover, hyperproliferative vascular diseases such asintimal smooth muscle cell hyperplasia, restenosis and vascularocclusion, particularly following biologically- or mechanically-mediatedvascular injury can be treated or prevented by the compounds of theinvention.

Other treatable conditions would include, but are not limited to,ischemic bowel diseases, inflammatory bowel diseases, necrotizingenterocolitis, intestinal lesions associated with thermal burns andleukotriene B4-mediated diseases; intestinal inflammations/allergiessuch as Coeliac diseases, proctitis, eosinophilic gastroenteritis,mastocytosis, Crohn's disease and ulcerative colitis; food-relatedallergic diseases which have symptomatic manifestation remote from thegastro-intestinal tract (e.g., migraine, rhinitis and eczema); renaldiseases such as interstitial nephritis, Goodpasture's syndrome,hemolytic-uremic syndrome and diabetic nephropathy; nervous diseasessuch as multiple myositis, Guillain-Barre syndrome, Meniere's disease,polyneuritis, multiple neuritis, mononeuritis and radiculopathy;endocrine diseases such as hyperthyroidism and Basedow's disease;hematic diseases such as pure red cell aplasia, aplastic anemia,hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmunehemolytic anemia, agranulocytosis, pernicious anemia, megaloblasticanemia and anerythroplasia; bone diseases such as osteoporosis;respiratory diseases such as sarcoidosis, fibroid lung and idiopathicinterstitial pneumonia; skin disease such as dermatomyositis, leukodermavulgaris, ichthyosis vulgaris, photoallergic sensitivity and cutaneous Tcell lymphoma; circulatory diseases such as arteriosclerosis,atherosclerosis, aortitis syndrome, polyarteritis nodosa andmyocardosis; collagen diseases such as scleroderma, Wegener's granulomaand Sjogren's syndrome; adiposis; eosinophilic fascitis; periodontaldisease such as lesions of gingiva, periodontium, alveolar bone andsubstantia ossea dentis; nephrotic syndrome such as glomerulonephritis;male pattern aleopecia or alopecia senilis by preventing epilation orproviding hair germination and/or promoting hair generation and hairgrowth; muscular dystrophy; Pyoderma and Sezary's syndrome; Addison'sdisease; active oxygen-mediated diseases, as for example organ injurysuch as ischemia reperfusion injury of organs (such as heart, liver,kidney and digestive tract) which occurs upon preservation,transplantation or ischemic disease (for example, thrombosis and cardiacinfraction); intestinal diseases such as endotoxin-shock,pseudomembranous colitis and colitis caused by drug or radiation; renaldiseases such as ischemic acute renal insufficiency and chronic renalinsufficiency; pulmonary diseases such as toxinosis caused bylung-oxygen or drug (for example, paracort and bleomycins), lung cancerand pulmonary emphysema; ocular diseases such as cataracta, siderosis,retinitis, pigmentosa, senile macular degeneration, vitreal scarring andcorneal alkali burn; dermatitis such as erythema multiforme, linear IgAballous dermatitis and cement dermatitis; and others such as gingivitis,periodontitis, sepsis, pancreatitis, diseases caused by environmentalpollution (for example, air pollution), aging, carcinogenis, metastasisof carcinoma and hypobaropathy; disease caused by histamine orleukotriene-C4 release; Behcet's disease such as intestinal-, vasculo-or neuro-Behcet's disease, and also Behcet's which affects the oralcavity, skin, eye, vulva, articulation, epididymis, lung, kidney and soon.

Furthermore, the compounds of the invention are useful for the treatmentand prevention of hepatic disease such as immunogenic diseases (forexample, chronic autoimmune liver diseases such as the group consistingof autoimmune hepatitis, primary biliary cirrhosis and sclerosingcholangitis), partial liver resection, acute liver necrosis (e.g.,necrosis caused by toxin, viral hepatitis, shock or anoxia), B-virushepatitis, non-A/non-B hepatitis, cirrhosis (such as alcoholiccirrhosis) and hepatic failure such as fulminant hepatic failure,late-onset hepatic failure and “acute-on chronic” liver failure (acuteliver failure on chronic liver diseases), and moreover are useful forvarious diseases because of their useful activity such as augmention ofchemotherapeutic effect, preventing or treating activity ofcytomegalovirus infection, particularly human cytomegalovirus (HCMV)infection, anti-inflammatory activity, and so on.

The compounds of the present invention may be used as vaccines to treatimmunosuppression in a subject. It is sometimes found that the antigenintroduced into the body for the acquisition of immunity from diseasealso acts as an immunosuppressive agent, and therefore, antibodies arenot produced by the body and immunity is not acquired. By introducing acompound of the present invention into the body as a vaccine, theundesired immunosuppression may be overcome and immunity acquired.

The compounds of the present invention may also find utility in thechemosensitization of drug resistant target cells. Cyclosporin A andFK-506 are known to be effective modulators of P-glycoprotein, asubstance which binds to and inhibits the action of anticancer drugs byinhibiting P-glycoprotein, as they are capable of increasing thesensitivity of multidrug resistant (MDR) cells to chemotherapeuticagents. It is believed that the compounds of the invention may likewisebe effective at overcoming resistance expressed to clinically usefulantitumour drugs such as 5-fluorouracil, cisplatin, methotrexate,vincristine, vinblastine and adriamycin, colchicine and vincristine.

Further, it has recently been shown that the steroid receptor-associatedheat shock proteins (hsp), hsp56 or hsp59, belong to the class ofimmunophilin proteins (see “HSP70 induction by cyclosporin A in culturedrat hepatocytes: effect of vitamin E succinate,” Andres, David et al.,Instituto de Bioqimica, Facultad de Farmacia, Universidad Complutense,Madrid, Spain. J. Hepatol. (2000) 33(4), 570-579; “Cyclosporin A Inducesan Atypical Heat Shock Response,” Paslaru, Liliana, et al., Biochem.Biophys. Res. Commun. (2000), 269(2), 464-469; “The cyclosporineA-binding immunophilin CyP-40 and the FK506-binding immunophilin hsp56bind to a common site on hsp90 and exist in independent cytosolicheterocomplexes with the untransformed glucocorticoid receptor,”Owens-Grillo, Janet K. et al., Med. Sch., Univ. Michigan, Ann Arbor,Mich. USA. J. Biol. Chem. (1995), 270(35), 20479-84). The ability of asteroid receptor 10 associated heat shock protein to bind theimmunosuppressive CsA suggests that the steroid receptor andimmunophilin signal transduction pathways are functionally interrelated.The combined treatment of compounds of the present invention and lowconcentrations of a steroid ligand (for e.g., progesterone,dexamethasone) result in a significant enhancement of target geneexpression over that seen in response to ligand alone. Thus, thecompounds of the present invention potentiate steroidmediatedtransactivation. Aqueous liquid compositions of the present inventionmay be particularly useful for the treatment and prevention of variousdiseases of the eye such as autoimmune diseases (including, for example,conical cornea, keratitis, dysophia epithelialis corneae, leukoma,Mooren's ulcer, scleritis and Graves' ophthalmopathy) and rejection ofcorneal transplantation.

Accordingly, the pharmaceutical compositions of the present inventioncomprise a therapeutically effective amount of a cyclosporin analog ofthe invention (e.g. those of the formulae delineated herein) incombination with a pharmaceutically acceptable carrier or excipient. Inparticular, compositions pertaining to the present invention are usefulfor treating a subject for immune-mediated organ or tissue allograftrejection, a graft-versus-host disease, an autoimmune disease, anobstructive airway disease, a hyperproliferative disease, or an ischemicor inflammatory intestinal or bowel disease.

The present invention also relates to method(s) of treatment of immunedisorders and inflammation or prevention of organ transplant rejectionin a subject by administering to the subject therapeutically effectiveamounts of the cyclosporin analogs of the present invention with orwithout the concurrent use of other drugs or pharmaceutically acceptableexcipients, as described throughout the present specification.

Yet a further embodiment of the present invention is a pharmaceuticalcomposition comprising any single compound delineated herein incombination with one or more HCV compounds known in the art, or apharmaceutically acceptable salt, ester, solvate, or prodrug thereof,with a pharmaceutically acceptable carrier or excipient.

It will be appreciated that reference herein to therapy and/or treatmentincludes, but is not limited to, prevention, retardation, prophylaxis,therapy and cure of the disease. It will further be appreciated thatreferences herein to treatment or prophylaxis of HCV infection includestreatment or prophylaxis of HCV-associated disease such as liverfibrosis, cirrhosis and hepatocellular carcinoma.

It will be further appreciated that the compounds of the presentinvention may contain one or more asymmetric carbon atoms and may existin racemic, diastereoisomeric, and optically active forms. It will stillbe appreciated that certain compounds of the present invention may existin different tautomeric forms. All tautomers are contemplated to bewithin the scope of the present invention.

It will be further appreciated that the compounds of the invention, ortheir pharmaceutically acceptable salts, stereoisomers, tautomers,prodrugs or salt of a prodrug thereof, can be administered as the soleactive pharmaceutical agent, or used in combination with one or moreagents to treat or prevent hepatitis C infections or the symptomsassociated with HCV infection. Other agents to be administered incombination with a compound or combination of compounds of the inventioninclude therapies for disease caused by HCV infection that suppressesHCV viral replication by direct or indirect mechanisms. These includeagents such as host immune modulators (for example, interferon-alpha,pegylated interferon-alpha, interferon-beta, interferon-gamma, CpGoligonucleotides and the like), or antiviral compounds that inhibit hostcellular functions such as inosine monophosphate dehydrogenase (forexample, ribavirin and the like). Also included are cytokines thatmodulate immune function. Also included are vaccines which comprise HCVantigens or antigen adjuvant combinations directed against HCV. Alsoincluded are agents that interact with host cellular components to blockviral protein synthesis by inhibiting the internal ribosome entry site(IRES) initiated translation step of HCV viral replication or to blockviral particle maturation and release with agents targeted toward theviroporin family of membrane proteins such as, for example, HCV P7 andthe like. Other agents to be administered in combination with a compoundof the present invention include any agent or combination of agents thatinhibit the replication of HCV by targeting proteins of the viral genomeinvolved in the viral replication. These agents include but are notlimited to other inhibitors of HCV RNA dependent RNA polymerase such as,for example, nucleoside type polymerase inhibitors described in WO01/90121(A2), or U.S. Pat. No. 6,348,587B1 or WO 01/60315 or WO 01/32153or non-nucleoside inhibitors such as, for example, benzimidazolepolymerase inhibitors described in EP1 162196A1 or WO 02/04425.

Accordingly, one aspect of the invention is directed to a method fortreating or preventing an infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment oneor more agents selected from the group consisting of a host immunemodulator and a second antiviral agent, or a combination thereof, with atherapeutically effective amount of a compound or combination ofcompounds of the invention, or a pharmaceutically acceptable salt,stereoisomer, tautomer, prodrug, salt of a prodrug, or combinationthereof. Examples of the host immune modulator are, but not limited to,interferon-alpha, pegylated-interferon-alpha, interferon-beta,interferon-gamma, a cytokine, a vaccine, and a vaccine comprising anantigen and an adjuvant, and said second antiviral agent inhibitsreplication of HCV either by inhibiting host cellular functionsassociated with viral replication or by targeting proteins of the viralgenome.

Further aspect of the invention is directed to a method of treating orpreventing infection caused by an RNA-containing virus comprisingco-administering to a patient in need of such treatment an agent orcombination of agents that treat or alleviate symptoms of HCV infectionincluding cirrhosis and inflammation of the liver, with atherapeutically effective amount of a compound or combination ofcompounds of the invention, or a pharmaceutically acceptable salt,stereoisomer, tautomer, prodrug, salt of a prodrug, or combinationthereof. Yet another aspect of the invention provides a method oftreating or preventing infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment oneor more agents that treat patients for disease caused by hepatitis B(HBV) infection, with a therapeutically effective amount of a compoundor a combination of compounds of the invention, or a pharmaceuticallyacceptable salt, stereoisomer, tautomer, prodrug, salt of a prodrug, orcombination thereof. An agent that treats patients for disease caused byhepatitis B (HBV) infection may be for example, but not limited thereto,L-deoxythymidine, adefovir, lamivudine or tenfovir, or any combinationthereof. Example of the RNA-containing virus includes, but not limitedto, hepatitis C virus (HCV).

Another aspect of the invention provides a method of treating orpreventing infection caused by an RNA-containing virus comprisingco-administering to a patient in need of such treatment one or moreagents that treat patients for disease caused by human immunodeficiencyvirus (HIV) infection, with a therapeutically effective amount of acompound or a combination of compounds of the invention, or apharmaceutically acceptable salt, stereoisomer, tautomer, prodrug, saltof a prodrug, or combination thereof. The agent that treats patients fordisease caused by human immunodeficiency virus (HIV) infection mayinclude, but is not limited thereto, ritonavir, lopinavir, indinavir,nelfinavir, saquinavir, amprenavir, atazanavir, tipranavir, TMC-114,fosamprenavir, zidovudine, lamivudine, didanosine, stavudine, tenofovir,zalcitabine, abacavir, efavirenz, nevirapine, delavirdine, TMC-125,L-870812, S-1360, enfuvirtide (T-20) or T-1249, or any combinationthereof. Example of the RNA-containing virus includes, but not limitedto, hepatitis C virus (HCV). In addition, the present invention providesthe use of a compound or a combination of compounds of the invention, ora therapeutically acceptable salt form, stereoisomer, or tautomer,prodrug, salt of a prodrug, or combination thereof, and one or moreagents selected from the group consisting of a host immune modulator anda second antiviral agent, or a combination thereof, to prepare amedicament for the treatment of an infection caused by an RNA-containingvirus in a patient, particularly hepatitis C virus. Examples of the hostimmune modulator are, but not limited to, interferon-alpha,pegylated-interferon-alpha, interferon-beta, interferon-gamma, acytokine, a vaccine, and a vaccine comprising an antigen and anadjuvant, and said second antiviral agent inhibits replication of HCVeither by inhibiting host cellular functions associated with viralreplication or by targeting proteins of the viral genome.

When used in the above or other treatments, combination of compound orcompounds of the invention, together with one or more agents as definedherein above, can be employed in pure form or, where such forms exist,in pharmaceutically acceptable salt form, prodrug, salt of a prodrug, orcombination thereof. Alternatively, such combination of therapeuticagents can be administered as a pharmaceutical composition containing atherapeutically effective amount of the compound or combination ofcompounds of interest, or their pharmaceutically acceptable salt form,prodrugs, or salts of the prodrug, in combination with one or moreagents as defined hereinabove, and a pharmaceutically acceptablecarrier. Such pharmaceutical compositions can be used for inhibiting thereplication of an RNA-containing virus, particularly Hepatitis C virus(HCV), by contacting said virus with said pharmaceutical composition. Inaddition, such compositions are useful for the treatment or preventionof an infection caused by an RNA-containing virus, particularlyHepatitis C virus (HCV).

Hence, further aspect of the invention is directed to a method oftreating or preventing infection caused by an RNA-containing virus,particularly a hepatitis C virus (HCV), comprising administering to apatient in need of such treatment a pharmaceutical compositioncomprising a compound or combination of compounds of the invention or apharmaceutically acceptable salt, stereoisomer, or tautomer, prodrug,salt of a prodrug, or combination thereof, one or more agents as definedhereinabove, and a pharmaceutically acceptable carrier.

When administered as a combination, the therapeutic agents can beformulated as separate compositions which are given at the same time orwithin a predetermined period of time, or the therapeutic agents can begiven as a single unit dosage form.

Antiviral agents contemplated for use in such combination therapyinclude agents (compounds or biologicals) that are effective to inhibitthe formation and/or replication of a virus in a mammal, including butnot limited to agents that interfere with either host or viralmechanisms necessary for the formation and/or replication of a virus ina mammal. Such agents can be selected from another anti-HCV agent; anHIV inhibitor; an HAV inhibitor; and an HBV inhibitor.

Other anti-HCV agents include those agents that are effective fordiminishing or preventing the progression of hepatitis C relatedsymptoms or disease. Such agents include but are not limited toimmunomodulatory agents, inhibitors of HCV NS3 protease, otherinhibitors of HCV polymerase, inhibitors of another target in the HCVlife cycle and other anti-HCV agents, including but not limited toribavirin, amantadine, levovirin and viramidine.

Immunomodulatory agents include those agents (compounds or biologicals)that are effective to enhance or potentiate the immune system responsein a mammal. Immunomodulatory agents include, but are not limited to,inosine monophosphate dehydrogenase inhibitors such as VX-497(merimepodib, Vertex Pharmaceuticals), class I interferons, class IIinterferons, consensus interferons, asialo-interferons pegylatedinterferons and conjugated interferons, including but not limited tointerferons conjugated with other proteins including but not limited tohuman albumin. Class I interferons are a group of interferons that allbind to receptor type I, including both naturally and syntheticallyproduced class I interferons, while class II interferons all bind toreceptor type II. Examples of class I interferons include, but are notlimited to, [alpha]-, [beta]-, [delta]-, [omega]-, and[tau]-interferons, while examples of class II interferons include, butare not limited to, [gamma]-interferons.

Inhibitors of HCV NS3 protease include agents (compounds or biologicals)that are effective to inhibit the function of HCV NS3 protease in amammal Inhibitors of HCV NS3 protease include, but are not limited to,those compounds described in WO 99/07733, WO 99/07734, WO 00/09558, WO00/09543, WO 00/59929, WO 03/064416, WO 03/064455, WO 03/064456, WO2004/030670, WO 2004/037855, WO 2004/039833, WO 2004/101602, WO2004/101605, WO 2004/103996, WO 2005/028501, WO 2005/070955, WO2006/000085, WO 2006/007700 and WO 2006/007708 (all by BoehringerIngelheim), WO 02/060926, WO 03/053349, WO03/099274, WO 03/099316, WO2004/032827, WO 2004/043339, WO 2004/094452, WO 2005/046712, WO2005/051410, WO 2005/054430 (all by BMS), WO 2004/072243, WO2004/093798, WO 2004/113365, WO 2005/010029 (all by Enanta), WO2005/037214 (Intermune) and WO 2005/051980 (Schering), and thecandidates identified as VX-950, ITMN-191 and SCH 503034.

Inhibitors of HCV polymerase include agents (compounds or biologicals)that are effective to inhibit the function of an HCV polymerase. Suchinhibitors include, but are not limited to, non-nucleoside andnucleoside inhibitors of HCV NS5B polymerase. Examples of inhibitors ofHCV polymerase include but are not limited to those compounds describedin: WO 02/04425, WO 03/007945, WO 03/010140, WO 03/010141, WO2004/064925, WO 2004/065367, WO 2005/080388 and WO 2006/007693 (all byBoehringer Ingelheim), WO 2005/049622 (Japan Tobacco), WO 2005/014543(Japan Tobacco), WO 2005/012288 (Genelabs), WO 2004/087714 (IRBM), WO03/101993 (Neogenesis), WO 03/026587 (BMS), WO 03/000254 (JapanTobacco), and WO 01/47883 (Japan Tobacco), and the clinical candidatesXTL-2125, HCV 796, R-1626 and NM 283.

Inhibitors of another target in the HCV life cycle include agents(compounds or biologicals) that are effective to inhibit the formationand/or replication of HCV other than by inhibiting the function of theHCV NS3 protease. Such agents may interfere with either host or HCVviral mechanisms necessary for the formation and/or replication of HCV.Inhibitors of another target in the HCV life cycle include, but are notlimited to, entry inhibitors, agents that inhibit a target selected froma helicase, a NS2/3 protease and an internal ribosome entry site (IRES)and agents that interfere with the function of other viral targetsincluding but not limited to an NS5A protein and an NS4B protein.

It can occur that a patient may be co-infected with hepatitis C virusand one or more other viruses, including but not limited to humanimmunodeficiency virus (HIV), hepatitis A virus (HAV) and hepatitis Bvirus (HBV). Thus also contemplated is combination therapy to treat suchco-infections by co-administering a compound according to the presentinvention with at least one of an HIV inhibitor, an HAV inhibitor and anHBV inhibitor.

DEFINITIONS

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “aryl,” as used herein, refers to a mono- or polycycliccarbocyclic ring system including, but not limited to, phenyl, naphthyl,tetrahydronaphthyl, indanyl, idenyl.

The term “heteroaryl,” as used herein, refers to a mono- or polycyclicaromatic radical having one or more ring atom selected from S, O and N;and the remaining ring atoms are carbon, wherein any N or S containedwithin the ring may be optionally oxidized. Heteroaryl includes, but isnot limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl,thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzooxazolyl, quinoxalinyl.

In accordance with the invention, any of the aryls, substituted aryls,heteroaryls and substituted heteroaryls described herein, can be anyaromatic group. Aromatic groups can be substituted or unsubstituted.

The terms “C₁-C₈ alkyl,” or “C₁-C₁₂ alkyl,” as used herein, refer tosaturated, straight- or branched-chain hydrocarbon radicals containingbetween one and eight, or one and twelve carbon atoms, respectively.Examples of C₁-C₈ alkyl radicals include, but are not limited to,methyl, ethyl, propyl, isopropyl, n-butyl, tent-butyl, neopentyl,n-hexyl, heptyl and octyl radicals; and examples of C₁-C₁₂ alkylradicals include, but are not limited to, ethyl, propyl, isopropyl,n-hexyl, octyl, decyl, dodecyl radicals.

The term “C₂-C₈ alkenyl,” as used herein, refer to straight- orbranched-chain hydrocarbon radicals containing from two to eight carbonatoms having at least one carbon-carbon double bond by the removal of asingle hydrogen atom. Alkenyl groups include, but are not limited to,for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl,heptenyl, octenyl, and the like.

The term “C₂-C₈ alkynyl,” as used herein, refer to straight- orbranched-chain hydrocarbon radicals containing from two to eight carbonatoms having at least one carbon-carbon triple bond by the removal of asingle hydrogen atom. Representative alkynyl groups include, but are notlimited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl,octynyl, and the like.

The term “C₃-C₈-cycloalkyl”, or “C₃-C₁₂-cycloalkyl,” as used herein,refers to a monocyclic or polycyclic saturated carbocyclic ringcompound. Examples of C₃-C₈-cycloalkyl include, but not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl andcyclooctyl; and examples of C₃-C₁₂-cycloalkyl include, but not limitedto, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,bicyclo[2.2.1]heptyl, and bicyclo[2.2.2]octyl.

The term “C₃-C₈ cycloalkenyl” or “C₃-C₁₂ cycloalkenyl” as used herein,refers to monocyclic or polycyclic carbocyclic ring compound having atleast one carbon-carbon double bond. Examples of C₃-C₈ cycloalkenylinclude, but not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like; and examples ofC₃-C₁₂ cycloalkenyl include, but not limited to, cyclopropenyl,cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl,and the like.

It is understood that any alkyl, alkenyl, alkynyl and cycloalkyl moietydescribed herein can also be an aliphatic group. Any cycloalkyl groupcan also be an alicyclic group. An “aliphatic” group is a non-aromaticmoiety that may contain any combination of carbon atoms, hydrogen atoms,halogen atoms, oxygen, nitrogen or other atoms, and optionally containone or more units of unsaturation, e.g., double and/or triple bonds. Analiphatic group may be straight chained, branched or cyclic andpreferably contains between about 1 and about 24 carbon atoms, moretypically between about 1 and about 12 carbon atoms. In addition toaliphatic hydrocarbon groups, aliphatic groups include, for example,polyalkoxyalkyls, such as polyalkylene glycols, polyamines, andpolyimines, for example. Such aliphatic groups may be furthersubstituted.

The term “alicyclic,” as used herein, denotes a monovalent group derivedfrom a monocyclic or bicyclic saturated carbocyclic ring compound by theremoval of a single hydrogen atom. Examples include, but not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptyl,and bicyclo[2.2.2]octyl. Such alicyclic groups may be furthersubstituted.

The terms “heterocyclic” or “heterocycloalkyl” can be usedinterchangeably and referred to a non-aromatic ring or a bi- ortri-cyclic group fused system, where (i) each ring system contains atleast one heteroatom independently selected from oxygen, sulfur andnitrogen, (ii) each ring system can be saturated or unsaturated (iii)the nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) thenitrogen heteroatom may optionally be quaternized, (v) any of the aboverings may be fused to an aromatic ring, and (vi) the remaining ringatoms are carbon atoms which may be optionally oxo-substituted.Representative heterocyclic groups include, but are not limited to,1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl,imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl,morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl,pyridazinonyl, and tetrahydrofuryl. Such heterocyclic groups may befurther substituted.

The term “substituted” refers to substitution by independent replacementof one, two, or three or more of the hydrogen atoms thereon withsubstituents including, but not limited to, —F, —Cl, —Br, —I, —OH,protected hydroxy, —NO₂, —CN, —N₃, —NH₂, protected amino, oxo, thioxo,—NH—C₁-C₁₂-alkyl, —NH—C₂-C₈-alkenyl, —NH—C₂-C₈-alkynyl,—NH—C₃-C₁₂-cycloalkyl, —NH-aryl, —NH-heteroaryl, —NH-heterocycloalkyl,-dialkylamino, -diarylamino, -diheteroarylamino, —O—C₁-C₁₂-alkyl,—O—C₂-C₈-alkenyl, —O—C₂-C₈-alkynyl, —O—C₃-C₁₂-cycloalkyl, —O-aryl,—O-heteroaryl, —O-heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl,—C(O)—C₂-C₈-alkenyl, —C(O)—C₂-C₈-alkynyl, —C(O)—C₃-C₁₂-cycloalkyl,—C(O)-aryl, —C(O)-heteroaryl, —C(O)-heterocycloalkyl, —CONH₂,—CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₈-alkenyl, —CONH—C₂-C₈-alkynyl,—CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl, —CONH-heteroaryl,—CONH-heterocycloalkyl, —OCO₂—C₁-C₁₂-alkyl, —OCO₂—C₂-C₈-alkenyl,—OCO₂—C₂-C₈-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl, —OCO₂-aryl,—OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl,—OCONH—C₂-C₈-alkenyl, —OCONH—C₂-C₈-alkynyl, —OCONH—C₃-C₁₂-cycloalkyl,—OCONH-aryl, —OCONH-heteroaryl, —OCONH-heterocycloalkyl,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₈-alkenyl, —NHC(O)—C₂-C₈-alkynyl,—NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl,—NHC(O)-heterocycloalkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₈-alkenyl,—NHCO₂—C₂-C₈-alkynyl, —NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl,—NHCO₂-heteroaryl, —NHCO₂-heterocycloalkyl, —NHC(O)NH₂,—NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₈-alkenyl,—NHC(O)NH—C₂-C₈-alkynyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₈-alkenyl,—NHC(S)NH—C₂-C₈-alkynyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl,—NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂,—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₈-alkenyl,—NHC(NH)NH—C₂-C₈-alkynyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl, —NHC(NH)NH-aryl,—NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₈-alkenyl, —NHC(NH)—C₂-C₈-alkynyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₈-alkenyl, —C(NH)NH—C₂-C₈-alkynyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₈-alkenyl,—S(O)—C₂-C₈-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₈-alkenyl, —SO₂NH—C₂-C₈-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₈-alkenyl, —NHSO₂—C₂-C₈-alkynyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₈-alkenyl, —S—C₂-C₈-alkynyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthiomethyl. It is understood that the aryls, heteroaryls, alkyls,and the like can be further substituted.

The term “halogen,” as used herein, refers to an atom selected fromfluorine, chlorine, bromine and iodine.

The term “hydroxy activating group”, as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxyl groupso that it will depart during synthetic procedures such as in asubstitution or an elimination reaction. Examples of hydroxyl activatinggroup include, but not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxy”, as used herein, refers to a hydroxy groupactivated with a hydroxyl activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxyl groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theart are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of hydroxyl protecting groups includebenzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, methoxycarbonyl, tert-butoxycarbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl,2-furfuryloxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl,trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl,2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, 1,1-dimethyl-2-propenyl,3-methyl-3-butenyl, allyl, benzyl, para-methoxybenzyldiphenylmethyl,triphenylmethyl(trityl), tetrahydrofuryl, methoxymethyl,methylthiomethyl, benzyloxymethyl, 2,2,2-trichloroethoxymethyl,2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, para-toluenesulfonyl,trimethylsilyl, triethylsilyl, triisopropylsilyl, and the like.Preferred hydroxyl protecting groups for the present invention areacetyl (Ac or —C(O)CH₃), benzoyl (Bz or —C(O)C₆H₅), and trimethylsilyl(TMS or —Si(CH₃)₃).

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups,for example.

The term “hydroxy prodrug group”, as used herein, refers to a promoietygroup which is known in the art to change the physicochemical, and hencethe biological properties of a parent drug in a transient manner bycovering or masking the hydroxy group. After said syntheticprocedure(s), the hydroxy prodrug group as described herein must becapable of reverting back to hydroxy group in vivo. Hydroxy prodruggroups as known in the art are described generally in Kenneth B. Sloan,Prodrugs, Topical and Ocular Drug Delivery, (Drugs and thePharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York(1992).

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the artare described generally in T. H. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York(1999). Examples of amino protecting groups include, but are not limitedto, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, andthe like.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

The term “protected amino,” as used herein, refers to an amino groupprotected with an amino protecting group as defined above.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such compounds are well known to those skilledin the art, and it will be obvious to those skilled in the art thatindividual solvents or mixtures thereof may be preferred for specificcompounds and reaction conditions, depending upon such factors as thesolubility of reagents, reactivity of reagents and preferred temperatureranges, for example. Further discussions of aprotic solvents may befound in organic chemistry textbooks or in specialized monographs, forexample: Organic Solvents Physical Properties and Methods ofPurification, 4th ed., edited by John A. Riddick et al., Vol. II, in theTechniques of Chemistry Series, John Wiley & Sons, NY, 1986.

The term “protic solvent’ as used herein, refers to a solvent that tendsto provide protons, such as an alcohol, for example, methanol, ethanol,propanol, isopropanol, butanol, t-butanol, and the like. Such solventsare well known to those skilled in the art, and it will be obvious tothose skilled in the art that individual solvents or mixtures thereofmay be preferred for specific compounds and reaction conditions,depending upon such factors as the solubility of reagents, reactivity ofreagents and preferred temperature ranges, for example. Furtherdiscussions of protogenic solvents may be found in organic chemistrytextbooks or in specialized monographs, for example: Organic SolventsPhysical Properties and Methods of Purification, 4th ed., edited by JohnA. Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, NY, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the Formula herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, 2^(nd) Ed. Wiley-VCH (1999); T. W. Greene and P. G. M.Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley andSons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The term “subject” as used herein refers to an animal. Preferably theanimal is a mammal. More preferably the mammal is a human. A subjectalso refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and may include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds, otherunsaturation, or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers or cis- and trans-isomers. Likewise, alltautomeric forms are also intended to be included. Tautomers may be incyclic or acyclic. The configuration of any carbon-carbon double bondappearing herein is selected for convenience only and is not intended todesignate a particular configuration unless the text so states; thus acarbon-carbon double bond or carbon-heteroatom double bond depictedarbitrarily herein as trans may be cis, trans, or a mixture of the twoin any proportion.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable saltsinclude, but are not limited to, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, maleic acid, tartaric acid,citric acid, succinic acid or malonic acid or by using other methodsused in the art such as ion exchange. Other pharmaceutically acceptablesalts include, but are not limited to, adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include, but are not limited to, formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds of the present invention which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals with undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, and effective for their intended use, aswell as the zwitterionic forms, where possible, of the compounds of thepresent invention. “Prodrug”, as used herein means a compound which isconvertible in vivo by metabolic means (e.g. by hydrolysis) to acompound of the invention. Various forms of prodrugs are known in theart, for example, as discussed in Bundgaard, (ed.), Design of Prodrugs,Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, vol. 4,Academic Press (1985); Krogsgaard-Larsen, et al., (ed). “Design andApplication of Prodrugs, Textbook of Drug Design and Development,Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug DeliverReviews, 8:1-38 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel DrugDelivery Systems, American Chemical Society (1975); and Bernard Testa &Joachim Mayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry,Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

The present invention also relates to solvates of the compounds ofFormula (I), for example hydrates.

This invention also encompasses pharmaceutical compositions containing,and methods of treating viral infections through administering,pharmaceutically acceptable prodrugs of compounds of the invention. Forexample, compounds of the invention having free amino, amido, hydroxy orcarboxylic groups can be converted into prodrugs. Prodrugs includecompounds wherein an amino acid residue, or a polypeptide chain of twoor more (e.g., two, three or four) amino acid residues is covalentlyjoined through an amide or ester bond to a free amino, hydroxy orcarboxylic acid group of compounds of the invention. The amino acidresidues include but are not limited to the 20 naturally occurring aminoacids commonly designated by three letter symbols and also includes4-hydroxyproline, hydroxylysine, demosine, isodemosine,3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,citrulline, homocysteine, homoserine, ornithine and methionine sulfone.Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. Freehydroxy groups may be derivatized using groups including but not limitedto hemisuccinates, phosphate esters, dimethylaminoacetates, andphosphoryloxymethyloxycarbonyls, as outlined in Advanced Drug DeliveryReviews, 1996, 19, 115. Carbamate prodrugs of hydroxy and amino groupsare also included, as are carbonate prodrugs, sulfonate esters andsulfate esters of hydroxy groups. Derivatization of hydroxy groups as(acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may bean alkyl ester, optionally substituted with groups including but notlimited to ether, amine and carboxylic acid functionalities, or wherethe acyl group is an amino acid ester as described above, are alsoencompassed. Prodrugs of this type are described in J. Med. Chem. 1996,39, 10. Free amines can also be derivatized as amides, sulfonamides orphosphonamides. All of these prodrug moieties may incorporate groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers or excipients.

As used herein, the term “pharmaceutically acceptable carrier orexcipient” means a non-toxic, inert solid, semi-solid or liquid filler,diluent, encapsulating material or formulation auxiliary of any type.Some examples of materials which can serve as pharmaceuticallyacceptable carriers are sugars such as lactose, glucose and sucrose;starches such as corn starch and potato starch; cellulose and itsderivatives such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; powdered tragacanth; malt; gelatin; talc; excipientssuch as cocoa butter and suppository waxes; oils such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; glycols such as propylene glycol; esters such as ethyloleate and ethyl laurate; agar; buffering agents such as magnesiumhydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffersolutions, as well as other non-toxic compatible lubricants such assodium lauryl sulfate and magnesium stearate, as well as coloringagents, releasing agents, coating agents, sweetening, flavoring andperfuming agents, preservatives and antioxidants can also be present inthe composition, according to the judgment of the formulator.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions, may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

For pulmonary delivery, a therapeutic composition of the invention isformulated and administered to the patient in solid or liquidparticulate form by direct administration e.g., inhalation into therespiratory system. Solid or liquid particulate forms of the activecompound prepared for practicing the present invention include particlesof respirable size: that is, particles of a size sufficiently small topass through the mouth and larynx upon inhalation and into the bronchiand alveoli of the lungs. Delivery of aerosolized therapeutics,particularly aerosolized antibiotics, is known in the art (see, forexample U.S. Pat. No. 5,767,068 to VanDevanter et al., U.S. Pat. No.5,508,269 to Smith et al., and WO 98/43650 by Montgomery, all of whichare incorporated herein by reference). A discussion of pulmonarydelivery of antibiotics is also found in U.S. Pat. No. 6,014,969,incorporated herein by reference.

According to the methods of treatment of the present invention, viralinfections, conditions are treated or prevented in a patient such as ahuman or another animal by administering to the patient atherapeutically effective amount of a compound of the invention, in suchamounts and for such time as is necessary to achieve the desired result.

By a “therapeutically effective amount” of a compound of the inventionis meant an amount of the compound which confers a therapeutic effect onthe treated subject, at a reasonable benefit/risk ratio applicable toany medical treatment. The therapeutic effect may be objective (i.e.,measurable by some test or marker) or subjective (i.e., subject gives anindication of or feels an effect). An effective amount of the compounddescribed above may range from about 0.1 mg/Kg to about 500 mg/Kg,preferably from about 1 to about 50 mg/Kg. Effective doses will alsovary depending on route of administration, as well as the possibility ofco-usage with other agents. It will be understood, however, that thetotal daily usage of the compounds and compositions of the presentinvention will be decided by the attending physician within the scope ofsound medical judgment. The specific therapeutically effective doselevel for any particular patient will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the activity of the specific compound employed; the specific compositionemployed; the age, body weight, general health, sex and diet of thepatient; the time of administration, route of administration, and rateof excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or contemporaneously with thespecific compound employed; and like factors well known in the medicalarts.

The total daily dose of the compounds of this invention administered toa human or other animal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

The compounds of the Formula described herein can, for example, beadministered by injection, intravenously, intraarterially, subdermally,intraperitoneally, intramuscularly, or subcutaneously; or orally,buccally, nasally, transmucosally, topically, in an ophthalmicpreparation, or by inhalation, with a dosage ranging from about 0.1 toabout 500 mg/kg of body weight, alternatively dosages between 1 mg and1000 mg/dose, every 4 to 120 hours, or according to the requirements ofthe particular drug. The methods herein contemplate administration of aneffective amount of compound or compound composition to achieve thedesired or stated effect. Typically, the pharmaceutical compositions ofthis invention will be administered from about 1 to about 6 times perday or alternatively, as a continuous infusion. Such administration canbe used as a chronic or acute therapy. The amount of active ingredientthat may be combined with pharmaceutically excipients or carriers toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. A typical preparation willcontain from about 5% to about 95% active compound (w/w). Alternatively,such preparations may contain from about 20% to about 80% activecompound.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Patients may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

When the compositions of this invention comprise a combination of acompound of the invention described herein and one or more additionaltherapeutic or prophylactic agents, both the compound and the additionalagent should be present at dosage levels of between about 1 to 100%, andmore preferably between about 5 to 95% of the dosage normallyadministered in a monotherapy regimen. The additional agents may beadministered separately, as part of a multiple dose regimen, from thecompounds of this invention. Alternatively, those agents may be part ofa single dosage form, mixed together with the compounds of thisinvention in a single composition.

The said “additional therapeutic or prophylactic agents” includes butnot limited to, immune therapies (eg. interferon), therapeutic vaccines,antifibrotic agents, anti-inflammatory agents such as corticosteroids orNSAIDs, bronchodilators such as beta-2 adrenergic agonists and xanthines(e.g. theophylline), mucolytic agents, anti-muscarinics,anti-leukotrienes, inhibitors of cell adhesion (e.g. ICAM antagonists),anti-oxidants (eg N-acetylcysteine), cytokine agonists, cytokineantagonists, lung surfactants and/or antimicrobial and anti-viral agents(eg ribavirin and amantidine). The compositions according to theinvention may also be used in combination with gene replacement therapy.

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one of ordinary skill in theart. All publications, patents, published patent applications, and otherreferences mentioned herein are hereby incorporated by reference intheir entirety.

Abbreviations

Abbreviations which may be used in the descriptions of the scheme andthe examples that follow are:

-   -   Ac for acetyl;    -   Boc₂O for di-tert-butyl-dicarbonate;    -   Boc for t-butoxycarbonyl;    -   Bz for benzoyl;    -   Bn for benzyl;    -   BocNHOH for tent-butyl N-hydroxycarbamate;    -   t-BuOK for potassium tert-butoxide;    -   BOP for (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium;    -   Hexafluorophosphate;    -   Brine for sodium chloride solution in water;    -   CDI for carbonyldiimidazole;    -   CH₂Cl₂ for dichloromethane;    -   CH₃ for methyl;    -   CH₃CN for acetonitrile;    -   Cs₂CO₃ for cesium carbonate;    -   dba for dibenzylidene acetone;    -   dppb for diphenylphosphino butane;    -   dppe for diphenylphosphino ethane;    -   DBU for 1,8-diazabicyclo[5.4.0]undec-7-ene;    -   DCC for N,N′-dicyclohexylcarbodiimide;    -   DEAD for diethylazodicarboxylate;    -   DIAD for diisopropyl azodicarboxylate;    -   DIPEA or (i-Pr)₂EtN for N,N,-diisopropylethyl amine;    -   Dess-Martin periodinane for        1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one;    -   DMAP for 4-dimethylaminopyridine;    -   DME for 1,2-dimethoxyethane;    -   DMF for N,N-dimethylformamide;    -   DMSO for dimethyl sulfoxide;    -   DPPA for diphenylphosphoryl azide;    -   EDC for N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide;    -   EDC HCl for N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide        hydrochloride;    -   EtOAc for ethyl acetate;    -   EtOH for ethanol;    -   Et₂O for diethyl ether;    -   HATU for        O-(7-azabenzotriazol-1-yl)-N,N,N′,N′,-tetramethyluronium;    -   Hexafluorophosphate;    -   HCl for hydrogen chloride;    -   HOBT for 1-hydroxybenzotriazole;    -   K₂CO₃ for potassium carbonate;    -   MeOH for methanol;    -   Ms for mesyl or —SO₂—CH₃;    -   Ms₂O for methanesulfonic anhydride or mesyl-anhydride;    -   NaHCO₃ for sodium bicarbonate or sodium hydrogen carbonate;    -   Na₂CO₃ sodium carbonate;    -   NaOH for sodium hydroxide;    -   Na₂SO₄ for sodium sulfate;    -   NaHSO₃ for sodium bisulfite or sodium hydrogen sulfite;    -   Na₂S₂O₃ for sodium thiosulfate;    -   NH₂NH₂ for hydrazine;    -   NH₄HCO₃ for ammonium bicarbonate;    -   NH₄Cl for ammonium chloride;    -   NMMO for N-methylmorpholine N-oxide;    -   NaIO₄ for sodium periodate;    -   OH for hydroxy;    -   OsO₄ for osmium tetroxide;    -   TEA or Et₃N for triethylamine;    -   TFA for trifluoroacetic acid;    -   THF for tetrahydrofuran;    -   TPP or PPh₃ for triphenylphosphine;    -   Ts for tosyl or —SO₂—C₆H₄CH₃;    -   Ts₂O for tolylsulfonic anhydride or tosyl-anhydride;    -   TsOH for p-tolylsulfonic acid;    -   Pd for palladium;    -   Ph for phenyl;    -   Pd₂(dba)₃ for tris(dibenzylideneacetone)dipalladium (0);    -   Pd(PPh₃)₄ for tetrakis(triphenylphosphine)palladium (0);    -   TBS for tent-butyl dimethylsilyl; or    -   TMS for trimethylsilyl;    -   TMSCl for trimethylsilyl chloride;    -   CsA for cyclosporinA.

Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared.

The novel cyclosporine analogues of the present invention are derivedfrom cyclosporine A. As shown in Scheme 1, a key intermediate of formula(1-3) was prepared by selective removal of amino acid in position four—N-methyl leucine of cyclosporinA (see Roland Wenger et al, “Syntheticroutes to NEtXaa⁴-cyclosporin A derivatives as potential anti-HIV Idrugs”, Tetrahedron Letters, 2000, 41, 7193, which is herebyincorporated by reference in its entirety). Thus, cyclosporin A wasreacted with acetic anhydride, optionally in the presence of pyridine orDMAP in CH₂Cl₂ to give acetylated intermediate (1-1), which was followedby selective cleavage of the amide bond between position three andposition four amino acid with trimethyloxonium tetrafluoroborate inCH₂Cl₂ to afford the intermediate (1-2). Edman degradation of (1-2) gavethe key intermediate (1-3).

Reduction of the compound of formula (1-3) with reducing agent such as,but not limited to NaBH₄ affords the compound of formula (1-4). Thereaction is carried out in a protic solvent such as, but not limited to,methanol, ethanol, isopropanol and tert-butanol or the mixture of twoprotic solvents. The reaction temperature can vary from 0° C. to about50° C. Protection of the amino group of the compound of formula (1-4)with Fmoc-Cl in the presence of organic base such as, but not limitedto, triethylamine, diisopropylethylamine, DBU, N-methylmorpholine andDMAP gives the compound of formula (1-5). The reaction is carried out inan aprotic solvent such as, but not limited to, CH₂Cl₂, DMF and THF. Thereaction temperature can vary from 0° C. to about 50° C. Furtherrearrangement of the compound of formula (1-5) in the presence of anacid, followed by acetyl protection gives the compound of formula (1-6).Suitable acids include, but are not limited to, methanesulfonic acid,toluenesulfonic acid, camphorsulfonic acid. The rearrangement reactionis carried out in a protic solvent such as, but not limited to,methanol, ethanol, isopropanol and tert-butanol. The acetyl protectionreaction is carried out in an aprotic solvent such as, but not limitedto, CH₂Cl₂, ClCH₂CH₂Cl, DMF and THF with acetic anhydride in thepresence of base. The suitable bases include, but are not limited to,triethylamine, diisopropylethylamine, DBU, N-methylmorpholine and DMAP.The compound of formula (1-6) is converted to the compound of formula(1-7) with sodium methoxide in methanol.

Scheme 2 illustrates a process of the invention for the preparation ofcompounds according to the invention. First the compound of formula(1-2) is coupled with a protected amino acid of the formula (2-1), whereW and R₃ is as previously defined to give the compound of formula (2-2).The coupling regent can be selected from, but not limited to DCC, EDC,di-isopropyl carbodiimide, BOP—Cl, PyBOP, PyAOP, TFFH and HATU. Suitablebases include, but are not limited to, triethylamine,diisopropylethylamine, DBU, N-methylmorpholine and DMAP. The couplingreaction is carried out in an aprotic solvent such as, but not limitedto, CH₂Cl₂, DMF and THF. The reaction temperature can vary from 0° C. toabout 50° C.

Then the compound of formula (2-2) is converted to the compound offormula (2-3) by NaBH₄ reduction in a protic solvent such as, but notlimited to, methanol, ethanol, isopropanol and tert-butanol or themixture of two protic solvents. Further rearrangement of the compound offormula (2-3) in the presence of an acid gives the compound of formula(2-4). Suitable acids include, but are not limited to, methanesulfonicacid, toluenesulfonic acid, camphorsulfonic acid. The rearrangementreaction is carried out in a protic solvent such as, but not limited to,methanol, ethanol, isopropanol and tert-butanol. The Boc group isdeprotected under this condition.

The compound of formula (2-4) is converted to the compound of formula(2-5) with sodium methoxide in methanol. The methyl ester of compound offormula (2-5) is converted to the corresponding acid compound of formula(2-6) via alkaline hydrolysis in protic solvents. Representative alkalicompounds include lithium hydroxide, sodium hydroxide, potassiumhydroxide, and the like. Suitable solvents include, but are not limitedto, methanol, ethanol, isopropanol, butanol, THF, 1,4-dioxane andmixtures thereof. The reaction temperature is preferably 0° to 35° C.

Compound of formula (2-7) is prepared by intramolecular amide formationreaction of the compound of formula (2-6). The reagent can be selectedfrom, but not limited to DCC, EDC, di-isopropyl carbodiimide, BOP—Cl,PyBOP, PyAOP, TFFH and HATU. Suitable bases include, but are not limitedto, triethylamine, diisopropylethylamine, DBU, N-methylmorpholine andDMAP. The coupling reaction is carried out in an aprotic solvent suchas, but not limited to, CH₂Cl₂, DMF and THF. The reaction temperaturecan vary from 0° C. to about 50° C.

Another alternative process for the preparation of the novelcyclosporinanalogues of the present invention is also illustrated inScheme 3.

First the compound of formula (1-7) is coupled with a protecteddipeptide of the formula (3-1), where W, R₃ and R₄ is as previouslydefined to give the compound of formula (3-2). The coupling regent canbe selected from, but not limited to DCC, EDC, di-isopropylcarbodiimide, BOP—Cl, PyBOP, PyAOP, TFFH and HATU. Suitable basesinclude, but are not limited to, triethylamine, diisopropylethylamine,DBU, N-methylmorpholine and DMAP. The coupling reaction is carried outin an aprotic solvent such as, but not limited to, CH₂Cl₂, DMF and THF.The reaction temperature can vary from 0° C. to about 50° C.

The protected dipeptides of formula (3-1) are prepared by the methoddescribed in Hu, T. and Panek, J. S.; J. Am. Chem. Soc. 2002, 124,11372.

The compound of formula (3-2) is converted to the compound of formula(3-3) by acidic Boc deprotection. The acid can be selected from, but notlimited to, TFA, HCl in dioxane, methanesulfonic acid. A more detaileddiscussion of the procedures, reagents and conditions for removingprotecting groups is described in literature, for example, by T. W.Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis”3^(rd) red., John Wiley & Son, Inc., 1999.

The methyl ester of compound of formula (3-3) is converted to thecorresponding acid compound of formula (3-4) via alkaline hydrolysis inprotic solvents. Representative alkali compounds include lithiumhydroxide, sodium hydroxide, potassium hydroxide, and the like. Suitablesolvents include, but are not limited to, methanol, ethanol,isopropanol, butanol, THF, 1,4-dioxane and mixtures there of. Thereaction temperature is preferably 0° to 35° C.

Compound of formula (3-5) is prepared by intramolecular amide formationreaction. The regent can be selected from, but not limited to DCC, EDC,di-isopropyl carbodiimide, BOP—Cl, PyBOP, PyAOP, TFFH and HATU. Suitablebases include, but are not limited to, triethylamine,diisopropylethylamine, DBU, N-methylmorpholine and DMAP. The couplingreaction is carried out in an aprotic solvent such as, but not limitedto, CH₂Cl₂, DMF and THF. The reaction temperature can vary from 0° C. toabout 50° C.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Although the invention has been described with respect to variouspreferred embodiments, it is not intended to be limited thereto, butrather those skilled in the art will recognize that variations andmodifications may be made therein which are within the spirit of theinvention and the scope of the appended claims.

Example 1 Compound of Formula IV: R₄=Isobutyl, W is Absent, and R₃=HStep 1a: Compound of Formula (1-1)

CsA (481 g, 0.4 mol) was dissolved in anhydrous CH₂Cl₂ (1.8 L). Aceticanhydride (163.3 g, 1.6 mol) was added followed by DMAP (48.86 g, 0.4mol) at room temperature under nitrogen. The reaction mixture wasstirred for 36 hrs. The reaction mixture was diluted with 6 L ofisopropyl acetate, followed by 8 L of water and stirred for 30 mins. Theorganic layer was separated and washed with saturated NaHCO₃ (4×6 L) andbrine (6 L). The organic phase was dried over Na₂SO₄ and concentrated.The resulted white foam was dried under vacuum to afford the compound offormula (1-1) (520 g, 95.5% HPLC purity).

MS (ESI): 1244.8 m/z (M+1).

Step 1b: Compound of Formula (1-2):

Compound of formula (1-1) (250 g, 0.2 mole) was dissolved in anhydrousCH₂Cl₂ (2 L). Trimethyloxoniumtetrafluoroborate (89.12 g, 0.6 mol) wasadded at 0° C. and the reaction mixture was stirred at room temperaturefor 20 hrs. Methanol and water (1:1 mixture, 2.5 L) was added via adropping funnel over 15 mins at 0° C. and then stirred at roomtemperature for 3 hrs. Reaction mixture was further diluted with 2 L ofCH₂Cl₂ and 2 L of water. The organic layer was separated and washed withsaturated Na₂CO₃ (2 L) and brine (2 L), and then dried over Na₂SO₄. Thesolvent was removed and the residue was purified on silica gel column toafford the compound of formula (1-2) (170 g, 92.5% HPLC purity).

MS (ESI): 1276.8 m/z (M+1).

Step 1c: Compound of Formula (1-3):

Compound of formula (1-2) (230 g, 0.18 mole) was dissolved in anhydrousTHF (1.5 L) and Phenyl thioisocyanate (24.35 g, 0.18 mole) was addedover 15 mins at 0° C. The reaction mixture was stirred at roomtemperature for 2 hrs and diluted with 1 L of water and 2.5 L of ethylacetate. The organic layer was separated and washed with brine (1 L),and then dried over Na₂SO₄, and concentrated. After dried under vacuumfor 24 hrs, the residue was dissolved in anhydrous CH₂Cl₂ (2.66 L). TFA(455 mL) was added at 0° C. over 30 mins and the reaction mixture wasstirred at room temperature for 4 hours. Reaction was quenched withsaturated Na₂CO₃ (3 L) at −15° C. The organic layer was separated andwashed with brine (3 L), and then dried over MgSO₄. Concentrated and theresidue was purified on silica gel column to afford the compound offormula (1-3) (130 g).

Step 1d: Compound of Formula (2-2):

To a solution of the compound of formula (1-3) (1.28 g, 1.0 mmol),Boc-D-proline (323 mg, 1.5 mmol) and DMAP (244 mg, 2.0 mmol) in DCM (10mL) was added HATU (0.76 g, 2 mmol) at 0° C. After stirred at roomtemperature for 14 hrs, the reaction mixture was quenched with 10%aqueous Na₂CO₃ and extracted with DCM. The organic layer was washed withsaturated KHSO₄ solution, brine and dried over anhydrous Na₂SO₄,filtered, concentrated, and purified by flash chromatography (ethylacetate/MeOH, 100-85%, v/v) to give the compound of formula (2-2) aswhite foam (1.1 g, 75% yield). ESI MS m/z=1473.84 [M+H]

Step 1e: Compound of Formula (2-3):

To a solution of the compound of formula (2-2) (1.1 g, 0.75 mmol) inmethanol (15 mL) was added NaBH₄ (500 mg, 10 portion in every 15 min.).After stirred at room temperature for 2 hrs, the reaction mixture wasquenched with 10% citric acid (50 ml) at 0° C. Extracted with ethylacetate (100 ml) and washed with 10% citric acid and brine, dried overanhydrous Na₂SO₄ and filtered. The solvent was removed and the residuewas purified on a silica gel column. (ethyl acetate/MeOH, 100-80%, v/v)to give the compound of formula (2-3) as white foam (1.0 g, 92% yield).ESI MS m/z=1445.88 [M+H]⁺.

Step 1f: Compound of Formula (2-4):

To a solution of the compound of formula (2-3) (1.0 g, 6.9 mmol) inmethanol (20 mL) was added methanesulfonic acid (2 ml). The reactionmixture was stirred at 50° C. for 3 hrs and the solvent was removed. Theresidue was diluted with ethyl acetate (50 ml) and added saturatedNa₂CO₃ (50 ml). The organic layer was washed with brine and dried overanhydrous Na2SO4. Concentrated and the residue was purified on silicagel column (DCM/MeOH, 100-70%, v/v) to give the compound of formula(2-4) as a white foam (0.8 g, 86% yield). ESI MS m/z=1345.82 [M+H]⁺.

Step 1g: Compound of Formula (2-6):

To a solution of the compound of formula (2-4) (0.8 g, 0.60 mmol) inmethanol (20 mL) was added NaOMe (300 mg) at 0° C. After stirred at roomtemperature for 8 hrs, NaOH (1N, 10 ml) was added and the mixture wasstirred at room temperature for 24 hrs. The reaction was quenched withsaturated NaHCO₃ (100 ml) and the mixture was extracted with ethylacetate (100 ml). The organic layer was washed with saturated NaHCO₃ andbrine. Concentrated to give the compound of formula (2-6) (350 mg, 49%),which was used for next step reaction without further purification.

Step 1h: Compound of Formula (2-7):

To a solution of HATU (152 mg, 0.4 mmol), DMAP (50 mg, 0.4 mmol) in DCM(200 mL) was added a solution of the compound of formula (2-6) (250 mg,0.2 mmol) in DCM (20 mL) in 2 hrs at room temperature. The solution wasstirred at room temperature for 16 hrs, concentrated, quenched withsaturated NaHCO₃ (100 ml). The organic layer was separated and driedover anhydrous Na₂SO₄. The solvent was removed and the residue waspurified on silica gel column to give the compound formula (2-7) aswhite foam (185 mg, 75% yield). ESI MS m/z=1228.90 [M+H]⁺.

Example 2 Compound of Formula IV: R₄=Isobutyl, W=O, and R₃=H

The compound of example 2 was prepared by using essential same procedureas the preparation of the compound of example 1.

Example 3 Compound of Formula IV: R₄=Isobutyl, W=O, and R₃=Me

A mixture of the compound of example 2 (25 mg), MeI (0.1 ml) and(t-Bu)₄NBr (20 mg) in DCM (1 ml) and 10N NaOH (1 ml) was stirred at roomtemperature for 48 hrs. The mixture was diluted with DCM (10 ml) andwashed with saturated NaHCO3. Dried over anhydrous Na2SO4 andconcentrated. The residue was purified on HPLC to give the compound ofexample 3 (20 mg, 79% yield). ESI MS m/z=1258.74 [M+H]⁺.

Example 4 Compound of Formula IV: R₄=Isobutyl, W=O, and R₃=Ac

The compound of example 4 was prepared by using essential same procedureas the preparation of the compound of example 1.

Example 5 Compound of Formula IV: R₄=Isobutyl, W=O, and R₃=allyl

The compound of example 5 was prepared by using essential same procedureas the preparation of the compound of example 3.

Example 6 Compound of Formula IV: R₄=Isobutyl, W=O, and R₃=—C(O)NH₂

To a solution of the compound of example 2 (25 mg, 0.02 mmol) andtriethyl amine (7 ul, 0.05 mmol) was added trichloroacetyl isocyanate(0.05 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 10min. and was added methanol (15 ml). The mixture was stirred at roomtemperature for 3 hrs. The solvent was removed and the residued waspurified on HPLC to give the compound of example 6 (18 mg, 70% yield).ESI MS m/z=1287.90 [M+H]⁺.

Example compounds 7-24 of the formula IV, wherein W, R₄ and R₃ aredelineated for each example in Table I are prepared from the compound ofexample 1 via the method delineated in example 3 and example 6.

TABLE I Ex- am- ple W R₄ R₃ 7 O

8 O

9 O

10 O

11 O

12 O

13 O

14 O

15 O

16 O

17 O

18 O

19 O

20 O

21 O

22 O

23 O

24 O

Example 25 Compound of Formula IV: R₄=Isopropyl, W=Absent, and R₃=H Step25a: Compound of Formula (1-3):

Compound of formula (1-2) (230 g, 0.18 mole) was dissolved in anhydrousTHF (1.5 L) and Phenyl thioisocyanate (24.35 g, 0.18 mole) was addedover 15 mins at 0° C. The reaction mixture was stirred at roomtemperature for 2 hrs and diluted with 1 L of water and 2.5 L of ethylacetate. The organic layer was separated and washed with brine (1 L),and then dried over Na₂SO₄, and concentrated. After dried under vacuumfor 24 hrs, the residue was dissolved in anhydrous CH₂Cl₂ (2.66 L). TFA(455 mL) was added at 0° C. over 30 mins and the reaction mixture wasstirred at room temperature for 4 hours. Reaction was quenched withsaturated Na₂CO₃ (3 L) at −15° C. The organic layer was separated andwashed with brine (3 L), and then dried over MgSO₄. Concentrated and theresidue was purified on silica gel column to afford the compound offormula (1-3) (130 g).

Step 25b: Compound of Formula (1-4):

Compound of formula (1-3) (61 g, 53 mmole) was dissolved in isopropanol(450 ml) and methanol (50 ml), and NaBH4 (9.0 g, 266 mmole) was addedduring 1 hrs 1 at 0° C. The reaction mixture was stirred at roomtemperature for 2 hrs. Ethyl acetate (50 ml) was added and the mixturewas stirred at room temperature for 30 min and then quenched with 1N HClat 0° C. The pH of mixture was adjusted to pH˜9 by adding saturatedNaHCO₃ and Na₂CO₃. Extracted with ethyl acetate and washed withsaturated NaHCO₃ and brine. Dried over Na₂SO₄ and the solvent wasremoved. The residue was dried on vacuum to give the compound of formula(1-4) (59.7 g)

Step 25c: Compound of Formula (1-5):

Compound of formula (1-4) (22.4 g, 20 mmole) was dissolved in DCM (200ml). FmocCl (4.92 g, 19 mmole) and DIPEA (5.17 g, 40 mmol) were added at0° C. The reaction mixture was stirred at 0° C. for 2 hrs. Diluted withDCM (500 ml) and washed with 10% citric acid, saturated NaHCO₃ andbrine. Dried over Na₂SO₄ and the solvent was removed. The residue waspurified on by silica gel column to give the compound of formula (1-5)(21 g).

Step 25d: Compound of Formula (1-6):

Compound of formula (1-5) (13.4 g, 10 mmole) was dissolved inisopropanol (100 ml). Methanesulfonic acid (100 mmole) was added at roomtemperature. The reaction mixture was stirred at 50° C. for 8 hrs. Thereaction mixture was condensed to ˜40 ml and was diluted with ethylacetate (500 ml) and quenched with saturated NaHCO₃. The pH of themixture was further adjusted to ˜9 by adding saturated Na₂CO₃. Organiclayer was separated and washed with brine. Dried over Na₂SO₄ and thesolvent was removed. The residue was dissolved in DCM (100 ml) and wasadded acetic anhydride (2.04 g, 20 mmol) followed by TEA (4.04 g, 40mmol). The mixture was stirred at room temperature for 3 hrs andquenched with saturated NaHCO3. The organic layer was separated andwashed with brine. Dried over Na₂SO₄ and concentrated. The residue waspurified on a silica gel column to give the compound of formula (1-6)(10 g).

Step 25e: Compound of Formula (1-7):

Compound of formula (1-6) (6.9 g, 5 mmole) was dissolved in methanol (50ml). NaOMe (2N in methanol, 25 ml) was added at room temperature. Thereaction mixture was stirred at room temperature for 18 hrs and quenchedwith saturated NaHCO₃. The pH of the mixture was further adjusted to ˜9by adding saturated Na₂CO₃. Organic layer was separated and washed withbrine. Dried over Na₂SO₄ and the solvent was removed. The residue waspurified on by silica gel column to give the compound of formula (1-7)(4.2 g).

Step 25f: Compound of Formula (3-2):

To a solution of the compound of formula (1-7) (1.04 g, 1.0 mmol), thecompound of formula (3-1), where R₄ is isopropyl, W is absent and R₃ isH (492 mg, 1.5 mmol) and DMAP (244 mg, 2.0 mmol) in DCM (20 mL) wasadded HATU (0.76 g, 2 mmol) at 0° C. After stirred at room temperaturefor 14 hrs, the reaction mixture was quenched with 10% aqueous Na₂CO₃and extracted with DCM. The organic layer was washed with saturatedKHSO₄ solution, brine and dried over anhydrous Na₂SO₄, filtered,concentrated, and purified by flash chromatography (ethyl acetate/MeOH,100-85%, v/v) to give the compound of formula (3-2) as white foam (1.15g, 85% yield). ESI MS m/z=1346.64 [M+H]⁺.

Step 25g: Compound of Formula (3-3):

A solution of the compound of formula (3-2) (1.08 g, 0.8 mmol) in DCM(10 ml) and TFA (5 ml) was stirred at room temperature for 2 hrs.Diluted with DCM (100 ml) and quenched with saturated Na2CO3 (100 ml).The organic layer was separated and washed with saturated Na₂CO₃ (100ml×2). Dried over anhydrous Na₂SO₄, filtered, concentrated to give crudecompound of formula (3-3) as white foam (0.94 g). ESI MS m/z=1246.74[M+H]⁺.

Step 25h: Compound of Formula (3-4):

To a solution of the compound of formula (3-3) (0.87 g, 0.7 mmol) in THF(20 ml) was added aqueous LiOH (0.5 N, 2.8 ml) at 0° C. The mixture wasstirred at 0° C. for 3 hrs and quenched with saturated NaHCO₃ (50 ml).Extracted with EtOAc (50 ml) and washed with brine (50 ml×2), dried overanhydrous Na₂SO₄, filtered, concentrated to give crude compound offormula (3-4) as white foam (0.73 g). ESI MS m/z=1232.64 [M+H]⁺.

Step 25i: Compound of Formula (3-5):

To a solution of HATU (380 mg, 1 mmol), DMAP (122 mg, 1 mmol) in DCM(200 mL) was added a solution of the compound of formula (3-4) (620 mg,0.5 mmol) in DCM (20 mL) in 2 hrs at room temperature. The solution wasstirred at room temperature for 16 hrs, concentrated, quenched withsaturated NaHCO₃ (100 ml). The organic layer was separated and driedover anhydrous Na₂SO₄. The solvent was removed and the residue waspurified on silica gel column to give the title compound of example 25as a white foam (480 mg, 80% yield). ESI MS m/z=1214.92 [M+H]⁺.

Example compounds 26-185 of the formula IV, wherein W, R₄ and R₃ aredelineated for each example in Table II, are prepared using the generalmethod set forth above for example 25.

TABLE II Example W R₄ R₃ 25 absent

H 26 O

H 27 O

Ac 28 O

allyl 29 O

30 O

31 O

32 O

33 O

34 O

35 O

36 O

37 O

38 O

39 O

40 O

41 O

42 O

43 O

44 O

45 O

46 O

47 O

48 O

H 49 O

H 50 O

Ac 51 O

allyl 52 O

53 O

54 O

55 O

56 O

57 O

58 O

59 O

60 O

61 O

62 O

63 O

64 O

65 O

66 O

67 O

68 O

69 O

70 O

71 O

H 72 O

H 73 O

Ac 74 O

allyl 75 O

76 O

77 O

78 O

79 O

80 O

81 O

82 O

83 O

84 O

85 O

86 O

87 O

88 O

89 O

90 O

91 O

92 O

93 O

94 O

H 95 O

H 96 O

Ac 97 O

allyl 98 O

99 O

100 O

101 O

102 O

103 O

104 O

105 O

106 O

107 O

108 O

109 O

110 O

111 O

112 O

113 O

114 O

115 O

116 O

117 O

H 118 O

H 119 O

Ac 120 O

allyl 121 O

122 O

123 O

124 O

125 O

126 O

127 O

128 O

129 O

130 O

131 O

132 O

133 O

134 O

135 O

136 O

137 O

138 O

139 O

140

H 141 O

H 142 O

Ac 143 O

allyl 144 O

145 O

146 O

147 O

148 O

149 O

150 O

151 O

152 O

153 O

154 O

155 O

156 O

157 O

158 O

159 O

160 O

161 O

162 O

163

H 164 O

H 165 O

Ac 166 O

allyl 167 O

168 O

169 O

170 O

171 O

172 O

173 O

174 O

175 O

176 O

177 O

178 O

179 O

180 O

181 O

182 O

183 O

184 O

185 O

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A compound represented by the formula (I);

or a pharmaceutically acceptable salt thereof, where: A is

 where, R₁ is selected from; a) R₁₁, where is selected from; 1)Hydrogen; 2) Deuterium; 3) C₁-C₈ alkyl; 4) Substituted C₁-C₈ alkyl; 5)C₂-C₈ alkenyl; 6) Substituted C₂-C₈ alkenyl; 7) C₂-C₈ alkynyl; 8)Substituted C₂-C₈ alkynyl; 9) C₃-C₁₂ cycloalkyl; 10) Substituted C₃-C₁₂cycloalkyl; 11) Aryl; 12) Substituted aryl; 13) Heterocycloalkyl; 14)Substituted heterocycloalkyl; 15) Heteroaryl; and 16) Substitutedheteroaryl; b) —C(O)OR₁₁, where R₁₁ is as previously defined; c)—C(O)R₁₁, where R₁₁ is as previously defined; d) —C(O)OCH₂—V—R₁₂, whereV is —O— or —S— and R₁₂ is selected from; a. C₁-C₈ alkyl; b. SubstitutedC₁-C₈ alkyl; c. C₂-C₈ alkenyl; d. Substituted C₂-C₈ alkenyl; e. C₂-C₈alkynyl; f. Substituted C₂-C₈ alkynyl; g. C₃-C₁₂ cycloalkyl; h.Substituted C₃-C₁₂ cycloalkyl; i. Aryl; j. Substituted aryl; k.Heterocycloalkyl; l. Substituted heterocycloalkyl; m. Heteroaryl; and n.Substituted heteroaryl; e) —C(O)N(R₁₃)(R₁₄), where R₁₃ and R₁₄ areindependently selected from R₁₁ and R₁₁ is as previously defined or R₁₃and R₁₄ taken together with the nitrogen atom to which they are attachedform a substituted or unsubstituted heterocycloalkyl; f) —C(O)SR₁₁,where R₁₁ is as previously defined; g) —C(S)OR₁₁, where R₁₁ is aspreviously defined; h) —C(O)OCH₂OC(O)R₁₂, where R₁₂ is as previouslydefined; i) —C(S)SR₁₁, where R₁₁ is as previously defined; and j) R₁₅,where R₁₅ is selected from; a. -M-R₁₁, where R₁₁ is as previouslydefined and M is selected from; i. C₁-C₈ alkyl; ii. Substituted C₁-C₈alkyl; iii. C₂-C₈ alkenyl; iv. Substituted C₂-C₈ alkenyl; v. C₂-C₈alkynyl; vi. Substituted C₂-C₈ alkynyl; vii. C₃-C₁₂ cycloalkyl; andviii. Substituted C₃-C₁₂ cycloalkyl; b. -M-NR₁₃R₁₄, where R₁₃, R₁₄ and Mare as previously defined; c. -M-S(O)_(m)R₁₁, where m=0, 1, or 2; M andR₁₁ are as previously defined; d. -M-OR₁₁, where M and R₁₁ are aspreviously defined; e. -M-C(O)R₁₁, where M and R₁₁ are as previouslydefined; f. -M-OC(O)R₁₂, where M and R₁₂ are as previously defined; g.-M—OC(O)OR₁₂, where M and R₁₂ are as previously defined; h.-M-NR₁₇C(O)R₁₂, where R₁₇ is R₁₁, M and R₁₂ are as previously defined;i. -MNR₁₇C(O)OR₁₂, where R₁₇, M and R₁₂ are as previously defined; j.-M-C(O)NR₁₃R₁₄, where R₁₃, M and R₁₄ are as previously defined; k.-M-C(O)N(R₁₇)—OR₁₁, where R₁₇, M and R₁₁ are as previously defined; l.-M-OC(O)NR₁₃R₁₄, where R₁₃, M and R₁₄ are as previously defined; m.-M-NR₁₇C(O)NR₁₃R₁₄, where M, R₁₃, R₁₇ and R₁₄ are as previously defined;n. -M-C(S)SR₁₁, where M and R₁₁ are as previously defined; o.-M-OC(S)SR₁₂, where M and R₁₂ are as previously defined; p.-M-NR₁₇C(O)SR₁₂, where M, R₁₇ and R₁₂ are as previously defined; q.-M-SC(O)NR₁₃R₁₄, where M, R₁₃ and R₁₄ are as previously defined; r.-M-CH═N—OR₁₁, where M and R₁₁ are as previously defined; and s.-M-CH═N—NR₁₃R₁₄, where M, R₁₃ and R₁₄ are as previously defined; B isethyl, 1-hydroxyethyl, isopropyl or n-propyl; X is OR₁ or SR₁, where R₁is as previously defined; W is absent, —O— or —S(O)_(m)—, where m=0, 1or 2; R₃ is R₁, where R₁ is as previously defined; R_(4N) is selectedfrom methyl, ethyl, allyl and propyl; and R₄ is—(CH₂)n₁-C(R₄₁)(R₄₂)—W₁—R₁, where n₁=0, 1 or 2; W₁ is absent, —O—, or—S(O)_(m)—, where m=0, 1 or 2; R₄₁ and R₄₂ are independently selectedfrom: hydrogen, methyl, ethyl, allyl, propyl or isopropyl; and R₁ is aspreviously defined.
 2. A compound according to claim 1 which isrepresented by the formula (II);

or a pharmaceutically acceptable salt thereof, wherein, R₃, R₄, R_(4N),and W are as defined in claim 1 and ═ represents a single bond or adouble bond.
 3. A compound according to claim 1 which is represented bythe formula (III);

or a pharmaceutically acceptable salt thereof, wherein, R₃, R₄ and W areas defined in claim
 1. 4. A compound according to claim 1 which isrepresented by the formula (IV);

or a pharmaceutically acceptable salt thereof, wherein R₃, R₄ and W areas defined in claim
 1. 5. A compound according to claim 1 which isselected from the group consisting of: Example 1: Compound of formulaIV: R₄=isobutyl, W is absent, and R₃=H; Example 2: Compound of formulaIV: R₄=isobutyl, W=O, and R₃=H; Example 3: Compound of formula IV:R₄=isobutyl, W=O, and R₃=Me; Example 4: Compound of formula IV:R₄=isobutyl, W=O, and R₃=Ac; Example 5: Compound of formula IV:R₄=isobutyl, W=O, and R₃=allyl; Example 6: Compound of formula IV:R₄=isobutyl, W=O, and R₃=—C(O)NH₂; and Example compounds 7-24 of theformula IV, wherein W, R₄ and R₃ are delineated for each example inTable I: TABLE I Ex- am- ple W R₄ R₃ 7 O

8 O

9 O

10 O

11 O

12 O

13 O

14 O

15 O

16 O

17 O

18 O

19 O

20 O

21 O

22 O

23 O

24 O

Example 25: Compound of formula IV: R₄=isopropyl, W=absent, and R₃=H:Example compounds 25-182 of the formula IV, wherein W, R₄ and R₃ aredelineated for each example in Table II: TABLE II Example W R₄ R₃ 25absent

H 26 O

H 27 O

Ac 28 O

allyl 29 O

30 O

31 O

32 O

33 O

34 O

35 O

36 O

37 O

38 O

39 O

40 O

41 O

42 O

43 O

44 O

45 O

46 O

47 O

48 absent

H 49 O

H 50 O

Ac 51 O

allyl 52 O

53 O

54 O

55 O

56 O

57 O

58 O

59 O

60 O

61 O

62 O

63 O

64 O

65 O

66 O

67 O

68 O

69 O

70 O

71 absent

H 72 O

H 73 O

Ac 74 O

allyl 75 O

76 O

77 O

78 O

79 O

80 O

81 O

82 O

83 O

84 O

85 O

86 O

87 O

88 O

89 O

90 O

91 O

92 O

93 O

94 absent

H 95 O

H 96 O

Ac 97 O

allyl 98 O

99 O

100 O

101 O

102 O

103 O

104 O

105 O

106 O

107 O

108 O

109 O

110 O

111 O

112 O

113 O

114 O

115 O

116 O

117 absent

H 118 O

H 119 O

Ac 120 O

allyl 121 O

122 O

123 O

124 O

125 O

126 O

127 O

128 O

129 O

130 O

131 O

132 O

133 O

134 O

135 O

136 O

137 O

138 O

139 O

140 absent

H 141 O

H 142 O

Ac 143 O

allyl 144 O

145 O

146 O

147 O

148 O

149 O

150 O

151 O

152 O

153 O

154 O

155 O

156 O

157 O

158 O

159 O

160 O

161 O

162 O

163 absent

H 164 O

H 165 O

Ac 166 O

allyl 167 O

168 O

169 O

170 O

171 O

172 O

173 O

174 O

175 O

176 O

177 O

178 O

179 O

180 O

181 O

182 O

183 O

184 O

185 O

or a pharmaceutically acceptable salt thereof.
 6. A pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof claim 1 or a pharmaceutically-acceptable salt, ester or prodrugthereof, in combination with a pharmaceutically acceptable carrier.
 7. Amethod of treating organ transplantation rejection in a subject, whichcomprises administering to said subject a therapeutically effectiveamount of the pharmaceutical composition of claim
 6. 8. A method oftreating an immune disorder in a subject, which comprises administeringto said subject a therapeutically effective amount of the pharmaceuticalcomposition of claim
 6. 9. The method of treating an immune disorder ina subject as set forth in claim 8, wherein said immune disorder isselected from the group consisting of psoriasis and eczema.
 10. Themethod of treating an immune disorder in a subject as set forth in claim9, wherein said administering is topical.
 11. The method of treating animmune disorder in a subject as set forth in claim 8, wherein saidimmune disorder is selected from the group consisting of: rheumatoidarthritis, inflammatory bowel disease, psoriasis, asthma, allergicrhinitis and chronic obstructive pulmonary disease.
 12. A method oftreating inflammatory or obstructive airways disease in a subject inneed of said treatment, comprising topically administering to saidsubject a therapeutically effective amount of the pharmaceuticalcomposition as defined in claim
 6. 13. The method of claim 12, whereinsaid topically administering is by inhalation.
 14. The method of claim12, wherein said airways disease is selected from the group consistingof asthma, allergic rhinitis, bronchitis, COPD including emphesema,chronic bronchitis and cystic fibrosis.
 15. A method of treating a viralinfection in a subject in need thereof, comprising administering to saidsubject a therapeutically-effective amount of a pharmaceuticalcomposition of claim
 6. 16. The method according to claim 15 whereinsaid viral infection is selected from HCV, HBV, HAV and HIV infection.17. The method of claim 16 further comprising coadministering one ormore additional anti-viral agents.
 18. The method of claim 17 whereinsaid additional anti-viral agents include: 1) current approved therapies(such as peg-interferon plus ribavirin), 2) viral-enzyme targetedcompounds, 3) viral-genome-targeted therapies (such as RNA interferenceor RNAi), and 4) immunomodulatory agents such ribavirin, interferon andToll-receptor agonists.
 19. The method of claim 17 further comprisingcoadministering with a CYP inhibitor.
 20. A process for the preparationof compounds of formula (I) as defined in claim 1.